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目的:探讨二十二碳六烯酸(DHA)对人肝癌细胞株Bel-7402增殖和凋亡的影响及其机制。方法:用不同浓度的DHA(0,25,50,100,200μmol/L)分别作用人肝癌Bel-7402细胞24,48,72 h后,用MTT法检测细胞的增殖情况、Western blot检测Bcl-2和Bax蛋白的表达,并分别用流式细胞仪、real-time PCR和caspase-3活性检测试剂盒检测上述梯度浓度的DHA作用Bel-7402细胞48 h后的凋亡情况、Bim基因表达和caspase-3活性。结果:不同浓度、不同时间DHA作用后,Bel-7402细胞增殖明显抑制,呈浓度和时间依赖性(均P<0.05);细胞Bax蛋白表达增加、Bcl-2蛋白表达降低,呈明显浓度依赖性(均P<0.05),但无明显时间依赖性(均P>0.05)。不同浓度DHA作用48 h后,Bel-7402细胞凋亡率、Bim基因表达和caspase-3的活性均明显增加,且均呈浓度依赖性(均P<0.05)。结论:DHA可抑制人肝癌Bel-7402细胞的增殖并诱导细胞凋亡,其机制可能与激活线粒体凋亡通路有关。
Objective: To investigate the effect of docosahexaenoic acid (DHA) on the proliferation and apoptosis of human hepatocellular carcinoma cell line Bel-7402 and its mechanism. METHODS: Human hepatocellular carcinoma Bel-7402 cells were treated with different concentrations of DHA (0, 25, 50, 100 and 200 μmol / L) for 24,48 and 72 h, respectively. Cell proliferation was measured by MTT assay. Western blot was used to detect Bcl- The apoptosis of Bel-7402 cells was detected by flow cytometry, real-time PCR and caspase-3 activity assay kit respectively. The expression of Bim and caspase-3 active. Results: The proliferation of Bel-7402 cells was inhibited in a concentration-and time-dependent manner (all P <0.05) after treated with DHA at different concentrations and different times. The expression of Bax protein and Bcl-2 protein were decreased in a concentration-dependent manner (All P <0.05), but no significant time-dependent (all P> 0.05). After treated with different concentrations of DHA for 48 h, the apoptosis rate, the expression of Bim and the activity of caspase-3 in Bel-7402 cells were significantly increased (all P <0.05). Conclusion: DHA can inhibit the proliferation of human hepatocellular carcinoma Bel-7402 cells and induce apoptosis, which may be related to the activation of mitochondrial apoptosis pathway.